cAMP (adenosine 3’5’-cyclic monophosphate) is a ubiquitous intracellular second messenger that mediates a diverse range of cellular processes in all organisms from bacteria to higher eukaryotes. It is converted from adenosine triphosphate (ATP) by adenylyl cyclases (ACs), and is inactivated by phosphodiesterases (PDEs) which catalyze its hydrolysis to 5’-AMP. In mammals, upon interaction with extracellular ligands, G protein coupled receptors (GPCRs) linked to Gαs activate the family of nine transmembrane ACs to increase intracellular cAMP. In contrast, GPCRs associated with Gαi/o inhibit the synthesis of cAMP by transmembrane ACs. cAMP binds and activates protein kinase A (PKA), the guanine nucleotide exchange factors Epac1/2, and cyclic nucleotide-gated ion channels. It can be released extracellularly where it is converted to adenosine by phosphodiesterases and nucleotidases.
cAMP Parameter Assay Kit
R&D Systems | Catalog # KGE002B
Key Product Details
Assay Length
Sample Type & Volume Required Per Well
Sensitivity
Assay Range
Product Summary for cAMP Parameter Assay Kit
Product Specifications
Measurement
Detection Method
Conjugate
Species
Specificity
Cross-reactivity
Interference
精度
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Saliva, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pmol/mL) | 22.6 | 68.7 | 116 | 21.7 | 70.4 | 119 |
| Standard Deviation | 1.37 | 2.55 | 2.54 | 2.2 | 6.0 | 7.3 |
| CV% | 6.0 | 3.7 | 2.2 | 10.3 | 8.6 | 6.1 |
Cell Lysates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pmol/mL) | 18.5 | 56.9 | 96.1 | 17.0 | 51.8 | 89.2 |
| Standard Deviation | 0.89 | 1.74 | 2.73 | 1.3 | 3.5 | 6.9 |
| CV% | 4.8 | 3.1 | 2.8 | 7.4 | 6.8 | 7.7 |
Recovery for cAMP Parameter Assay Kit
The recovery of cAMP spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 104 | 86-119 |
| Cell Lysis Buffer (n=1) | 91 | 87-94 |
| EDTA Plasma (n=4) | 105 | 92-120 |
| Heparin Plasma (n=4) | 98 | 85-111 |
| Saliva (n=4) | 97 | 80-117 |
| Serum (n=4) | 103 | 97-114 |
| Urine (n=6) | 100 | 91-119 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of cAMP were serially diluted with the appropriate diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for cAMP Parameter Assay Kit
Multi-species cAMP ELISA Cell Lysis Buffer 5 (1X) Standard Curve
Multi-species cAMP ELISA Calibrator Diluent RD5-55 Standard Curve
Preparation and Storage
Shipping
Stability & Storage
Background: cAMP
Long Name
Alternate Names
Additional cAMP Products
Product Documents for cAMP Parameter Assay Kit
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for cAMP Parameter Assay Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for cAMP Parameter Assay Kit
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FAQs for cAMP Parameter Assay Kit
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Q: The cAMP Parameter Assay Kit, Catalog # KGE002B, provides a cell lysate preparation protocol. Can the cell freezing times be varied when using this protocol?
A: Cell lysates were prepared using the lysate preparation protocol as described. While our scientists have not tested alternate preparation protocols, alteration of the freezing time for cells (including overnight freezing) and using more than two freeze-thaws is not expected to alter the assay results. To be safe, user can determine this with some preliminary testing.